Adiponectin upregulates ferritin heavy chain in skeletal muscle cells.
OBJECTIVE
Adiponectin is an adipocyte-derived protein that acts to reduce insulin resistance in the liver and muscle and also inhibits atherosclerosis. Although adiponectin reportedly enhances AMP-activated protein kinase and inhibits tumor necrosis factor-alpha action downstream from the adiponectin signal, the precise physiological mechanisms by which adiponectin acts on skeletal muscles remain unknown.


METHODS
We treated murine primary skeletal muscle cells with recombinant full-length human adiponectin for 12 h and searched, using two-dimensional electrophoresis, for proteins upregulated more than threefold by adiponectin compared with untreated cells.


RESULTS
We found one protein that was increased 6.3-fold with adiponectin incubation. MALDI-TOF (matrix-assisted laser desorption/ionization-top of flight) mass spectrometric analysis identified this protein as ferritin heavy chain (FHC). When murine primary skeletal muscle cells were treated with adiponectin, IkappaB-alpha phosphorylation was observed, suggesting that adiponectin stimulates nuclear factor (NF)-kappaB activity. In addition, FHC upregulation by adiponectin was inhibited by NF-kappaB inhibitors. These results suggest NF-kappaB activation to be involved in FHC upregulation by adiponectin. Other NF-kappaB target genes, manganese superoxide dismutase (MnSOD) and inducible nitric oxide synthase (iNOS), were also increased by adiponectin treatment. We performed a reactive oxygen species (ROS) assay using CM-H(2)DCFDA fluorescence and found that ROS-reducing effects of adiponectin were abrogated by FHC or MnSOD small-interfering RNA induction.


CONCLUSIONS
We have demonstrated that adiponectin upregulates FHC in murine skeletal muscle tissues, suggesting that FHC elevation might partially explain how adiponectin protects against oxidative stress in skeletal muscles.
Ikegami
Yuichi
Y
Department of Endocrinology and Diabetes, School of Medicine, Saitama Medical University, Saitama, Japan.
Inukai
Kouichi
K
Imai
Kenta
K
Sakamoto
Yasushi
Y
Katagiri
Hideki
H
Kurihara
Susumu
S
Awata
Takuya
T
Katayama
Shigehiro
S
D052242:Adiponectin / Q000494:pharmacology*; D000818:Animals; D001052:Apoferritins / Q000378:metabolism*; D015153:Blotting, Western; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D015870:Gene Expression / Q000187:drug effects; D051379:Mice; D018482:Muscle, Skeletal / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism
D052242:Adiponectin; D016328:NF-kappa B; D017382:Reactive Oxygen Species; D001052:Apoferritins; D052247:Nitric Oxide Synthase Type II; D013482:Superoxide Dismutase